Anti-SRC Family (phospho Y418) 抗体 [EP503Y] (ab40660)

製品の概要

  • 製品名Anti-SRC Family (phospho Y418) antibody [EP503Y]
    SRC Family 一次抗体 製品一覧
  • 製品の詳細
    Rabbit Monoclonal [EP503Y] to SRC Family (phospho Y418)
  • 特異性ab40660 recognises Lyn phosphorylation on Tyrosine 396. BLAST analysis show 100% homology with Hck (pY410) and Lck (pY393), and 92% homology with Src (pY418), Fyn (pY419) and Yes (pY425) Therefore, athe antibody may cross-react with these proteins.
  • アプリケーション適用あり: WB, Flow Cyt, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide within Human SRC Family aa 350-450 (phospho Y418). The exact sequence is proprietary.
    Database link: P07948
    (Peptide available as ab202645)

  • ポジティブ・コントロール
    • WB: HeLa and C6 cell lysates treated with pervanadate. ICC/IF: HeLa cells treated with pervanadate. Flow Cyt: HeLa cells treated with pervanadate.
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.
    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    This protein is known to be similar in amino acid sequence to HCK (P08631), LCK (P06239), FYN (P06241), YES1 (P07947), and SRC (P12931). Therefore, cross-reactivity with these homologous proteins may be observed. We would be happy to provide immunogen alignment information upon request.

    Alternative versions available:
    Anti-Lyn (phospho Y396) antibody (Alexa Fluor® 594) [EP503Y] (ab206347)

     

製品の特性

  • 製品の状態Liquid
  • 保存方法Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • バッファーPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • Concentration information loading...
  • ポリ/モノモノクローナル
  • クローン名EP503Y
  • アイソタイプIgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab40660 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB 1/1000 - 1/10000. Detects a band of approximately 56 kDa (predicted molecular weight: 61 kDa).Can be blocked with Lyn (phospho Y396) peptide (ab202645).

Induction of cells with Pervanadate: Prepare 500mM stock Sodium Orthovanadate in PBS. To make 100mM (100x) Pervanadate Na3VO4 right before experiment, add 400ul 500mM Sodium Orthovanadate to 1560ul H2O and then 40ul H2O2, vortex and incubate at room RT for 5-10min. Cells are serum-starved overnight, and then treated with freshly made Pervanadate (100x, final concentration 1mM) for 20min at 37°C.

Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.

ターゲット情報

  • 関連性SRC Family Negative Regulatory Phosphorylation Site belongs to the protein kinase superfamily, Tyr protein kinase family, SRC subfamily. It contains one protein kinase domain, one SH2 domain and one SH3 domain. It may serve as part of a signaling pathway coupling the Fc receptor to the activation of the respiratory burst and may also contribute to neutrophil migration and may regulate the degranulation process of neutrophils. SRC Family Negative Regulatory Phosphorylation Site is expressed predominantly in cells of the myeloid and B lymphoid lineages. There are two named isoforms.
  • 細胞内局在Isoform 1: Membrane; Lipid anchor. Isoform 2: Membrane; Lipid anchor. Cytoplasm.
  • 参照データベース
  • 別名
    • ASV antibody
    • Protooncogene SRC, Rous sarcoma antibody
    • SRC antibody
    • SRC1 antibody

Anti-SRC Family (phospho Y418) antibody [EP503Y] 画像

  • All lanes : Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660) at 1/2000 dilution (purified)

    Lane 1 : Untreated HeLa whole cell lysate
    Lane 2 : Pervanadate treated HeLa whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 61 kDa
    Observed band size : 56-59 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 seconds

    Blocking and dilution buffer: 5% BSA/TBST.

  • All lanes : Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660) at 1/1000 dilution (purified)

    Lane 1 : Untreated C6 whole cell lysate
    Lane 2 : Pervanadate treated C6 whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 61 kDa
    Observed band size : 56-59 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 second

    Blocking and dilution buffer: 5% BSA/TBST.

  • All lanes : Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660) at 1/1000 dilution (purified)

    Lane 1 : Pervanadate treated Jurkat cell lysate with Lyn non-phospho peptide
    Lane 2 : Pervanadate treated Jurkat cell lysate with Lyn (pY396) phospho peptide
    Lane 3 : Pervanadate treated Jurkat cell lysate with Lck non-phospho peptide
    Lane 4 : Pervanadate treated Jurkat cell lysate with Lck (pY393) phospho peptide
    Lane 5 : Pervanadate treated Jurkat cell lysate with Src non-phospho peptide
    Lane 6 : Pervanadate treated Jurkat cell lysate with Src (pY418) phospho peptide

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 61 kDa
    Observed band size : 56-59 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 minutes

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells (untreated and treated with 1mM pervanadate for 15 minutes) labelling SRC Family (phospho Y418) with purified ab40660 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/250) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • ICC/IF image of unpurified ab40660 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab40660, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Flow Cytometry analysis of HeLa cells (treated with 1mM pervanadate for 30 minutes) labelling SRC Family (phospho Y418) with purified ab40660 at 1/80 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody.

    Black - Isotype control, rabbit monoclonal IgG with pervanade treated HeLa cells.

    Blue - Unlabelled control, pervanadate treated HeLa cells without incubation with primary and secondary antibodies.

    Red - Pervandate treated HeLa cells with ab40660.

    Green - Untreated HeLa cells labelled with ab40660.

  • Direct ELISA antibody dose-response curve using unpurified ab40660 at 0-1000 ng/ml. Antigen concentration of 1000 ng/mL. An alkaline phosphatase-conjugated goat anti-rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

Anti-SRC Family (phospho Y418) antibody [EP503Y] (ab40660) 使用論文

This product has been referenced in:
  • Chiricozzi E  et al. Direct interaction, instrumental for signaling processes, between LacCer and Lyn in the lipid rafts of neutrophil-like cells. J Lipid Res 56:129-41 (2015). Read more (PubMed: 25418321) »
  • Nagao T  et al. Proliferation and survival signaling from both Jak2-V617F and Lyn involving GSK3 and mTOR/p70S6K/4EBP1 in PVTL-1 cell line newly established from acute myeloid leukemia transformed from polycythemia vera. PLoS One 9:e84746 (2014). Read more (PubMed: 24404189) »

See all 12 Publications for this product

Product Wall

Application Western blot
Loading amount 60 µg
Gel Running Conditions Reduced Denaturing (10% PAGE Gel)
Sample Human Cell lysate - whole cell (Hela cells)
Specification Hela cells
Treatment 100ng/ml IL6 for 15 minutes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 Feb 12 2015

The difference between lyn a and lyn b is that lyn b does not contain amino acids 23-43 of lyn a. Since this antibody recognises a region around tyrosine 396, ab40660 can detect both lyn a and lyn b therefore it is very probable that this doublet band ...

Read More

Thank you for your enquiry regarding ab40660.
Based on the sequence homology, it is unlikely that this antibody would cross react with the other tyrosines of Lyn. However, it might cross-react with LCK, HCK and possibly Src when phosphorylated at ...

Read More

ab33906 lot YH090207C - 0.3260 mg/ml

ab33862 lot YH082410D - 0.6380 mg/ml

ab32429 lot YG080503 - 0.0790 mg/ml

ab40660 lot YI070802CS - 0.1030 mg/ml

ab32445 lot YI072804DS - 0.0250 mg/ml

Thank you for your enquiry.

I can confirm that we have a dedicated team at Abcam who review the literature in order to find publications that have used our products. In this case, the following publication was found, showing use of this anti...

Read More

Thank you for your enquiry.

These 2 antibodies have lot specific concentrations. If you could please forward me the lot numbers for ab40660 and ab33914 I'd be happy to enquire about the concentrations.

Thanks for the additional info...

Read More

Thank you for contacting Abcam.

We added the reactivity with mouse based on the following reference:

Kim SW et al. Rational combined targeting of phosphodiesterase 4B and SYK in DLBCL. Blood 113:6153-60 (2009).



...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"